Reaction mass of Trisodium tris(D-gluconato(2-)-κO1,κO2)ferrate(3-) and Sodium chloride

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25-31 July 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: EU method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: OECD Guideline no. 439: In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of Trisodium tris(D-gluconato(2-)-κO1,κO2)ferrate(3-) and Sodium chloride

EC Number:

946-138-3

Molecular formula:

C18H30FeNa3O21 + NaCl

IUPAC Name:

Reaction mass of Trisodium tris(D-gluconato(2-)-κO1,κO2)ferrate(3-) and Sodium chloride

Test material form:

solid: particulate/powder

Details on test material:

Appearance Yellow powder Visual
Assay 76.4% NMR
NaCl 19.3% CZE/titration
water 2.7% Karl Fisher titration
pH 9.5 (1% aqueous solution)
Expiry date: November 2017

Specific details on test material used for the study:

Yellow powder
Purity: 76.4%
Batch no.: FC-C 11988
Expiry date: 1 November 2017

In vitro test system

Test system:

human skin model

Vehicle:

unchanged (no vehicle)

Details on test system:

TEST SITE
- Area of exposure: human epidermis model
- % coverage: 0.38 cm2

REMOVAL OF TEST SUBSTANCE
- Washing (if done): phosphate buffered saline
- Time after start of exposure: 15 minutes

POST INCUBATION PERIOD
- 42 hours

SCORING SYSTEM:
- After a 42 hour incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment. Cell viability was calculated for each tissue as a percentage of the mean of the negative control tissues.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

The test was performed on a total of 3 tissues per test item together with negative and positive controls. The skin was moistened with 5 µl Milli-Q water (Millipore Corp., Bedford, Mass., USA) to ensure close contact of the test item to the tissue and the solid test item (15.18 to 20.30 mg) was added into 12-well plates on top of the skin tissues. Three tissues were treated with 25 µl PBS (negative control) and 3 tissues with 25 µl 5% SDS (positive control) respectively. The positive control was re-spread after 7 minutes contact time.

Duration of treatment / exposure:

Exposure:15 minutes
Post incubation period: 42 hours

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

other: other: percentage viability

Value:

116

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other:

Remarks:

Basis: other: percentage of control. Time point: 15 minutes. (migrated information)

Any other information on results incl. tables

The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with Iron trigluconic acid, trisodium salt compared to the negative control tissues was 116%. Since the mean relative tissue viability for Iron trigluconic acid, trisodium salt was above 50%, Iron trigluconic acid, trisodium salt is considered to be non-irritant.

The positive control had a mean cell viability after 15 ± 0.5 minutes exposure of 5.7%. The absolute mean OD₅₇₀of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was ≤ 18%, indicating that the test system functioned properly.

Applicant's summary and conclusion

Interpretation of results:

other: non-irritant

Remarks:

Migrated information

Conclusions:

It is concluded that the test substance is non-irritant in the in vitro skin irritation test.

Executive summary:

The objective of this study was to evaluate Iron trigluconic acid, trisodium salt for its ability to induce skin irritation on a human three dimensional epidermal model (EPISKIN Small model (EPISKIN-SM^TM)). The possible skin irritation potential of Iron trigluconic acid, trisodium salt was tested through topical application for 15 minutes.

The study procedures described in this report were based on the most recent OECD and EC guidelines.

Batch FC-C 11988 of the test item was a yellow powder. Skin tissue was moistened with 5 µl of Milli-Q water and at least 10 mg of the test item was applied directly on top of the skin tissue for 15 ± 0.5 minutes. After a 42 hour post-incubation period, determination of the cytotoxic (irritancy) effect was performed. Cytotoxicity is expressed as the reduction of mitochondrial dehydrogenase activity measured by formazan production from MTT at the end of the treatment.

Skin irritation is expressed as the remaining cell viability after exposure to the test item. The relative mean tissue viability obtained after 15 ± 0.5 minutes treatment with Iron trigluconic acid, trisodium salt compared to the negative control tissues was 116%. Since the mean relative tissue viability for Iron trigluconic acid, trisodium salt was above 50% after 15 ± 0.5 minutes treatment, Iron trigluconic acid, trisodium salt is considered to be non-irritant.

The positive control had a mean cell viability of 5.7% after 15 ± 0.5 minutes exposure. The absolute mean OD₅₇₀(optical density at 570 nm) of the negative control tissues was within the laboratory historical control data range. The standard deviation value of the percentage viability of three tissues treated identically was ≤ 18%, indicating that the test system functioned properly.

In conclusion, Iron trigluconic acid, trisodium salt is non-irritant in the in vitro skin irritation test under the experimental conditions described in this report andshould not be classified according to the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) of the United Nations.